Moving toward point-of-care surgery in Ukraine: testing an ultra-portable operating room in an active war zone.

PURPOSE: In conflict zones, providers may have to decide between delaying time-sensitive surgeries or performing operative interventions in the field, potentially subjecting patients to significant infection risks. We conducted a single-arm crossover study to assess the feasibility of using an ultraportable operating room (U-OR) for surgical procedures on a porcine cadaver abdominal traumatic injury model in an active war zone. METHODS: We enrolled participants from an ASSET-type course designed to train Ukrainian surgeons before deployment to active conflict zones. They performed three standardized consecutive abdominal surgical procedures (liver, kidney, and small bowel injury repair) with and without the U-OR. Primary outcomes included surgical procedure completion rate, procedure time, and airborne particle count at the start of surgery. Secondary survey-based outcomes assessed surgery task load index (SURG-TLX) and perceived operative factors. RESULTS: Fourteen surgeons performed 76 surgical procedures (38 with the U-OR, 38 without the U-OR). The completion rate for each surgical procedure was 100% in both groups. While the procedure time for the liver injury repair did not differ significantly between the two groups, the use of the U-OR was associated with a longer time for kidney (155 vs. 56 s, p = 0.002), and small bowel (220 vs. 103 s, p = 0.004) injury repair. The average airborne particle count within the U-OR was substantially lower compared to outside the U-OR (6,753,852 vs. 232,282 n/m3, p < 0.001). There was no statistically significant difference in SURG-TLX for procedures performed with and without the U-OR. CONCLUSION: The use of the U-OR did not affect the procedure completion rate or SURG-TLX. However, there was a marked difference in airborne particle counts between inside and outside the U-OR during surgery. These preliminary findings indicate the potential feasibility of using a U-OR to perform abdominal damage-control surgical procedures in austere settings.

Changes in dairy cows' behaviour, health, and production after transition from tied to loose housing.

BACKGROUND: Transition of dairy cows from a tied to a loose housing system may affect their behaviour, health and production. Such housing system changes have become more frequent in Estonia but knowledge is lacking on how cows adapt to a new system. The aim of this study was to evaluate how cows' behaviour, milk production and composition, and different aspects of their health changed after transition from tied to loose housing. RESULTS: A herd of 400 dairy cows was moved to a new system on the same farm, so that effects of transport were not confounding factors. Behavioural observations were made for approximately 4 months following transition. Milk production data were recorded from 12 months before to 12 months after transition. Examination for skin alterations and cleanliness, as well as body condition scoring were carried out before transition, and thereafter monthly throughout the study. Significant effects on behaviour were observed just after the transition, with increases in the behaviour indicative of poor welfare, such as vocalisation and aggression, and decreases in those indicative of a good state of welfare, such as ruminating, resting and grooming. These effects were of short duration, with most returning to a steady state after the first week. Milk production declined already before the transition but fell significantly after transition, and this fall lasted longer in older cows. Likewise, somatic cell counts were higher in all cows following transition, but older cows were affected significantly more than cows in the first lactation. The frequency of lameness and skin alterations increased on average after transition. Body condition scores fell after transition but recovered by the second month. Therefore, there were adverse effects on the behaviour, health and production of the dairy cows transferred, although, apart from older cows, of short duration. CONCLUSION: The transition from tied to loose housing first had negative impacts on the welfare of the cows, although by the tenth day the behavioural indicators had returned to normal values. Impacts were more severe in higher parity cows, indicating that the change was more of a challenge for older cows. The findings of this study suggest that animals' behaviour and health should be more carefully observed within about 2 weeks after transition. It is quite likely that more and more farmers in Estonia and elsewhere will recognize the benefits of keeping their dairy cattle in loose housing, aimed at improving animal welfare and the value of the production chain.

Hip Abductor Deficiency after Total Hip Arthroplasty: Diagnostic and Therapeutic Methods. / Glutealinsuffizienz nach Hüfttotalendoprothese: Diagnostik und Therapieverfahren.

Abductor deficiency after total hip arthroplasty is a severe complication with functional limitations and a significant reduction in the patient's quality of life. Common causes are degenerative ruptures or approach-related iatrogenic damage to the gluteus medius and minimus muscle and the inferior gluteal nerve, fractures of the greater trochanter and incorrect reconstruction of leg length and femoroacetabular offset. With a standardised diagnosis consisting of a clinical examination, conventional X-ray and MRI, the causes of the functional problems can often be reliably determined. Therapy of abductor deficiency is challenging for both patients and physicians and is often tedious. However, with a clear diagnostic and therapeutic algorithm and straightforward patient education, good treatment results can be achieved even in this challenging condition. Conservative therapy with eccentric stretching and muscle strengthening are the basis of the treatment. In cases of progression of complaints despite intensive conservative treatment, various anatomical and extra-anatomical surgical reconstruction methods are available to relieve pain and improve function. Anatomical reconstruction of the gluteal tendon insertion is an option in cases of low-grade fatty infiltration and moderate retraction of the gluteal muscles. In situations with advanced degenerative changes in the gluteus medius and minimus muscles and an intact gluteus maximus muscle, transfer of the anterior portion of the gluteus maximus according to Whiteside is an option. For high-grade defects of the soft tissue, there is also the option of an isolated or combined transfer of the vastus lateralis muscle.

IL-15 Superagonist NAI in BCG-Unresponsive Non-Muscle-Invasive Bladder Cancer.

BACKGROUND: Patients with Bacillus Calmette­Guérin (BCG)­unresponsive non­muscle-invasive bladder cancer (NMIBC) have limited treatment options. The immune cell­activating interleukin-15 (IL-15) superagonist Nogapendekin alfa inbakicept (NAI), also known as N-803, may act synergistically with BCG to elicit durable complete responses (CRs) in this patient population. METHODS: In this open-label, multicenter study, patients with BCG-unresponsive bladder carcinoma in situ (CIS) with or without Ta/T1 papillary disease were treated with intravesical NAI plus BCG (cohort A) or NAI alone (cohort C). Patients with BCG-unresponsive high-grade Ta/T1 papillary NMIBC also received NAI plus BCG (cohort B). The primary end point was the incidence of CR at the 3- or 6-month assessment visit for cohorts A and C, and the disease-free survival (DFS) rate at 12 months for cohort B. Durability, cystectomy avoidance, progression-free survival, disease-specific survival (DSS), and overall survival were secondary end points for cohort A. RESULTS: In cohort A, CR was achieved in 58 (71%) of 82 patients (95% confidence interval [CI]=59.6 to 80.3; median follow-up, 23.9 months), with a median duration of 26.6 months (95% CI=9.9 months to [upper bound not reached]). At 24 months in patients with CR, the Kaplan­Meier estimated probability of avoiding cystectomy and of DSS was 89.2% and 100%, respectively. In cohort B (n=72), the Kaplan­Meier estimated DFS rate was 55.4% (95% CI=42.0% to 66.8%) at 12 months, with median DFS of 19.3 months (95% CI=7.4 months to [upper bound not reached]). Most treatment-emergent adverse events for patients receiving BCG plus NAI were grade 1 to 2 (86%); three grade 3 immune-related treatment-emergent adverse events occurred. CONCLUSIONS: In patients with BCG-unresponsive bladder carcinoma in situ and papillary NMIBC treated with BCG and the novel agent NAI, CRs were achieved with a persistence of effect, cystectomy avoidance, and 100% bladder cancer­specific survival at 24 months. The study is ongoing, with an estimated target enrollment of 200 participants (Funded by ImmunityBio.)

Knowledge, attitude and perceptions of medical students towards mental health in a university in Uganda.

BACKGROUND: The prevalence of mental illness among medical students is high. A gap remains on what knowledge should be given to improve the attitudes and perceptions towards mental health. Despite the vast body of literature globally, no study has been conducted in Uganda to assess the levels of knowledge, attitude, and perception among medical students in Uganda. OBJECTIVE: To determine the level of knowledge, attitude, and perception and their associated factors among medical students in Uganda. METHODS: A cross-sectional study was done among 259 undergraduate medical students in a public university capturing information on knowledge, attitude, and perception towards mental health. Linear regression analysis was used to determine the factors associated with knowledge, attitude, and perception. RESULTS: About 77.72% had high knowledge, 49.29% had positive attitudes, and 46.92% had good perceptions of mental health. There was a significant positive relationship between attitude and perceptions towards mental illness. At multilevel analysis, being in year 4 increased the level of knowledge (ß = 1.50 [95% confidence interval (CI) = 0.46-2.54], p = 0.005) while a positive history of mental illness worsened perceptions towards mental illness (ß = -4.23 [95% CI = -7.44-1.03], p = 0.010). CONCLUSION: Medical students have a high level of knowledge about mental illness but the majority had poor attitudes and perceptions of mental illness. Exposure to psychiatry knowledge about mental illness in year four increased students' knowledge while prior experience with mental illness conditions was associated with poorer perceptions. The information present in this study can be used by policymakers and future researchers to design future studies and interventions to improve knowledge, perceptions, and attitudes especially among students who have a history of mental illness. Improvements in knowledge, attitude, and perception may improve the mental health services for the future patients of these medical students.

Perspectives on mental health services for medical students at a Ugandan medical school.

BACKGROUND: University-based mental health services for medical students remain a challenge, particularly in low-income countries, due to poor service availability. Prior studies have explored the availability of mental health services in high-income countries but little is known about mental health services in countries in sub-Saharan Africa, such as Uganda. Medical students are at a higher risk of developing mental health challenges during their course of study as compared with other students. Thus, there is a need for well-structured mental health services for this group of students. The aim of this study was to explore perspectives on mental health services for medical students at a public University in Uganda. METHODS: This was a qualitative study where key informant interviews were conducted among purposively selected university administrators (n = 4), student leaders (n = 4), and mental health employees of the university (n = 3), three groups responsible for the mental well-being of medical students at a public university in Uganda. Interviews were audio-recorded, transcribed, and thematically analyzed to identify relevant themes. RESULTS: The working experience of university administrators and mental health providers was between eight months to 20 years, while student leaders had studied at the university for over four years. We identified five broad themes: (1) Burden of medical school: A curriculum of trauma, (2) Negative coping mechanisms and the problem of blame, (3) The promise of services: Mixed Messages, (4) A broken mental health system for students, and (5) Barriers to mental health services. CONCLUSION: Distinguishing between psychological distress that is anticipated because of the subject matter in learning medicine and identifying those students that are suffering from untreated psychiatric disorders is an important conceptual task for universities. This can be done through offering education about mental health and well-being for administrators, giving arm's length support for students, and a proactive, not reactive, approach to mental health. There is also a need to redesign the medical curriculum to change the medical education culture through pedagogical considerations of how trauma informs the learning and the mental health of students.

Measured weight loss as a precursor to cancer diagnosis: retrospective cohort analysis of 43 302 primary care patients.

BACKGROUND: Unexpected weight loss is a presenting feature of cancer in primary care. Data from primary care are lacking to quantify how much weight loss over what period should trigger further investigation for cancer. This research aimed to quantify cancer diagnosis rates associated with measured weight change in people attending primary care. METHODS: Retrospective cohort study of primary care electronic health records data linked to the Surveillance, Epidemiology, and End Results cancer registry (Integrated healthcare delivery system in Washington State, United States). Multivariable Cox regression incorporating time varying covariates using splines to model non-linear associations (age, percentage weight change, and weight change interval). Fifty thousand randomly selected patients aged 40 years and over followed for up to 9 years (1 January 2006 to 31 December 2014). Outcome measures are hazard ratios (95% confidence intervals) to quantify the association between percentage weight change and cancer diagnosis for all cancers combined, individual cancer sites and stages; percentage risk of cancer diagnosis within 6 months of the end of each weight change episode; and the positive predictive value for cancer diagnosis. RESULTS: There were 43 302 included in the analysis after exclusions. Over 287 858 patient-years of follow-up, including 24 272 (56.1%) females, 23 980 (55.4%) aged 40 to 59 years, 15 113 (34.9%) 60 to 79 years, and 4209 (9.7%) aged 80 years and over. Adjusted hazard ratios (95% confidence interval) for cancer diagnosis in a 60 years old ranged from 1.04 (1.02 to 1.05, P < 0.001) for 1% weight loss to 1.44 (1.23 to 1.68, P < 0.001) for 10%. An independent linear association was observed between percentage weight loss and increasing cancer risk. The absolute risk of cancer diagnosis increased with increasing age (up to 85 years) and as the weight change measurement interval decreased (<1 year). The positive predictive value for a cancer diagnosis within 1 year of ≥5% measured weight loss in a 60 to 69 years old was 3.41% (1.57% to 6.37%) in men and 3.47% (1.68% to 6.29%) in women. The risk of cancer diagnosis was significantly increased for pancreatic, myeloma, gastro-oesophageal, colorectal, breast, stage II and IV cancers. CONCLUSIONS: Weight loss is a sign of undiagnosed cancer regardless of the interval over which it occurs. Guidelines should resist giving an arbitrary cut-off for the interval of weight loss and focus on the percentage of weight loss and the patient's age. Future studies should focus on the association between diagnostic evaluation of weight change and risk of cancer mortality.

The Effect of High-Altitude Acclimatisation on Ultra-Short Heart Rate Variability.

Introduction: High-altitude (HA) exposure affects heart rate variability (HRV) and has been inconsistently linked to acute mountain sickness (AMS). The influence of increasing HA exposure on ultra-short HRV and its relationship to gold standard HRV measures at HA has not been examined. Methods: This was a prospective observational study of adults aged ≥ 18 years undertaking a HA trek in the Dhaulagiri region of the Himalayas. Cardiac inter-beat-intervals were obtained from a 10-s recording of supra-systolic blood pressure (Uscom BP+ device) immediately followed by 300 s single lead ECG recording (CheckMyHeart device). HRV was measured using the RMSSD (root mean square of successive differences of NN intervals) at sea level (SL) in the United Kingdom and at 3,619, 4,600, and 5,140 m at HA. Oxygen saturations (SpO2) were measured using finger-based pulse oximetry. The level of agreement between the 10 and 300 s RMSSD values were examined using a modified Bland-Altman relative-difference analysis. Results: Overall, 89 participants aged 32.2 ± 8.8 years (range 18-56) were included of which 70.8% were men. HA exposure (SL vs. 3,619 m) was associated with an initial increase in both 10 s (45.0 [31.0-82.0]) vs. 58.0 [33.0-119.0] ms) and 300 s (45.67 [33.24-70.32] vs. 56.48 [36.98-102.0] ms) in RMSSD. Thereafter at 4,600 and 5,140 m both 10 and 300 s RMSSD values were significantly lower than SL. From a total of 317 paired HRV measures the 10 and 300 s RMSSD measures were moderately correlated (Spearman r = 0.66; 95% CI: 0.59-0.72; p < 0.0001). The median difference (bias) in RMSSD values (300 s - 10 s) was -2.3 ms with a lower and upper limit of agreement of -107.5 and 88.61 ms, respectively with no differences with altitude. Overall, 293/317 (92.4%) of all paired HRV values fell within the 95% CI limits of agreement. Neither HRV method was predictive of AMS. Conclusion: Increasing HA affects ultra-short HRV in a similar manner to gold-standard 300 s. Ultra-short HRV has a moderate agreement with 300 s measurements. HRV did not predict AMS.

Black Babies Matter.

Despite dramatic advancements in neonatal intensive care since the 1960s, African-American infants still have more than a two-fold higher first-year mortality rate than non-Latinx White infants. Our essay examines the impact of upstream factors closely linked to the historical and contemporary context of structural racism in the United States on the African-American women's birth outcome disadvantage. In the process, we propose a paradigm to address the racial health inequity in adverse birth outcome by considering the interplay of racism and social class.

Use of Tribulus terrestris and Lepidium meyenii extract in rats: reproductive, biochemical and body parameters.

The oral administration of Tribulus terrestris and Lepidium meyenii extracts on reproductive, biochemical and body parameters was evaluated in rats. Thirty-six male Wistar rats weighting 210 ± 18 g were divided into six experimental groups (n = 6). Each group received, daily for 28 days, different solutions: T. terrestris (100 mg/kg), L. meyenii (1 g/kg) and T. terrestris at doses of 75, 50 and 25 mg/kg combined with L. meyenii at doses of 0.25, 0.5 and 0.75 g/kg, respectively, and distilled water (control). T. terrestris increased (p < 0.05) the serum testosterone, regardless of dose. Combined use of the extracts increased (p < 0.05) the diameter of the epididymal duct and epididymis lumen. The combinations of T. terrestris (75 and 50 mg/kg) with L. meyenii increased (p < 0.05) the sperm concentration. There were no differences (p > 0.05) in the other semen characteristics; relative weight of organs; and serum levels of urea, creatinine, alanine and aspartate transaminase, gamma glutamyl transferase, cholesterol, triglycerides, glucose, follicle-stimulating hormone and luteinizing hormone. No histopathological changes were observed (p > 0.05). It is concluded that the association of T. terrestris and L. meyenii has positive effects on serum testosterone, sperm concentration and epididymal morphology, with no evidence of effects in the testis, liver, spleen and kidneys.

Oxidized GAPDH transfers S-glutathionylation to a nuclear protein Sirtuin-1 leading to apoptosis.

AIMS: S-glutathionylation is a reversible oxidative modification of protein cysteines that plays a critical role in redox signaling. Glutaredoxin-1 (Glrx), a glutathione-specific thioltransferase, removes protein S-glutathionylation. Glrx, though a cytosolic protein, can activate a nuclear protein Sirtuin-1 (SirT1) by removing its S-glutathionylation. Glrx ablation causes metabolic abnormalities and promotes controlled cell death and fibrosis in mice. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH), a key enzyme of glycolysis, is sensitive to oxidative modifications and involved in apoptotic signaling via the SirT1/p53 pathway in the nucleus. We aimed to elucidate the extent to which S-glutathionylation of GAPDH and glutaredoxin-1 contribute to GAPDH/SirT1/p53 apoptosis pathway. RESULTS: Exposure of HEK 293T cells to hydrogen peroxide (H2O2) caused rapid S-glutathionylation and nuclear translocation of GAPDH. Nuclear GAPDH peaked 10-15 min after the addition of H2O2. Overexpression of Glrx or redox dead mutant GAPDH inhibited S-glutathionylation and nuclear translocation. Nuclear GAPDH formed a protein complex with SirT1 and exchanged S-glutathionylation to SirT1 and inhibited its deacetylase activity. Inactivated SirT1 remained stably bound to acetylated-p53 and initiated apoptotic signaling resulting in cleavage of caspase-3. We observed similar effects in human primary aortic endothelial cells suggesting the GAPDH/SirT1/p53 pathway as a common apoptotic mechanism. CONCLUSIONS: Abundant GAPDH with its highly reactive-cysteine thiolate may function as a cytoplasmic rheostat to sense oxidative stress. S-glutathionylation of GAPDH may relay the signal to the nucleus where GAPDH trans-glutathionylates nuclear proteins such as SirT1 to initiate apoptosis. Glrx reverses GAPDH S-glutathionylation and prevents its nuclear translocation and cytoplasmic-nuclear redox signaling leading to apoptosis. Our data suggest that trans-glutathionylation is a critical step in apoptotic signaling and a potential mechanism that cytosolic Glrx controls nuclear transcription factors.

Characterization of a Sex-Determining Region and Its Genomic Context via Statistical Estimates of Haplotype Frequencies in Daughters and Sons Sequenced in Pools.

Sex chromosomes are generally derived from a pair of autosomes that have acquired a locus controlling sex. Sex chromosomes may evolve reduced recombination around this locus and undergo a long process of molecular divergence. At that point, the original loci controlling sex may be difficult to pinpoint. This difficulty has affected many model species from mammals to birds to flies, which present highly diverged sex chromosomes. Identifying sex-controlling loci is easier in species with molecularly similar sex chromosomes. Here we aimed at pinpointing the sex-determining region (SDR) of Armadillidium vulgare, a terrestrial isopod with female heterogamety (ZW females and ZZ males) and whose sex chromosomes appear to show low genetic divergence. To locate the SDR, we assessed single-nucleotide polymorphism (SNP) allele frequencies in F1 daughters and sons sequenced in pools (pool-seq) in several families. We developed a Bayesian method that uses the SNP genotypes of individually sequenced parents and pool-seq data from F1 siblings to estimate the genetic distance between a given genomic region (contig) and the SDR. This allowed us to assign more than 43 Mb of contigs to sex chromosomes, and to demonstrate extensive recombination and very low divergence between these chromosomes. By taking advantage of multiple F1 families, we delineated a very short genomic region (∼65 kb) that presented no evidence of recombination with the SDR. In this short genomic region, the comparison of sequencing depths between sexes highlighted female-specific genes that have undergone recent duplication, and which may be involved in sex determination in A. vulgare.

Why does racial inequity in health persist?

Attempts over decades have failed to eliminate the glaring inequity in birth outcomes between Americans of different races. We propose broadening the approach to dealing with racial health inequity by considering the interplay of race and class in the politics of the United States. This approach, combined with a grasp of the historical roots of race relations in North America, could hold the promise of improving our country's abysmal showing in international comparisons of population health indicators, including birth outcomes.

Validation of an mRNA-based Urine Test for the Detection of Bladder Cancer in Patients with Haematuria.

BACKGROUND: In patients with haematuria, a fast, noninvasive test with high sensitivity (SN) and negative predictive value (NPV), which is able to detect or exclude bladder cancer (BC), is needed. A newly developed urine assay, Xpert Bladder Cancer Detection (Xpert), measures five mRNA targets (ABL1, CRH, IGF2, UPK1B, and ANXA10) that are frequently overexpressed in BC. OBJECTIVE: To validate the performance of Xpert in patients with haematuria. DESIGN, SETTING, AND PARTICIPANTS: Voided precystoscopy urine specimens were prospectively collected at 22 sites from patients without prior BC undergoing cystoscopy for haematuria. Xpert, cytology, and UroVysion procedures were performed. Technical validation was performed and specificity (SP) was determined in patients without BC. OUTCOME MEASUREMENTS AND STATISTICAL ANALYSIS: Test characteristics were calculated based on cystoscopy and histology results, and compared between Xpert, cytology, and UroVysion. RESULTS AND LIMITATIONS: We included 828 patients (mean age 64.5 yr, 467 males, 401 never smoked). Xpert had an SN of 78% (95% confidence interval [CI]: 66-87) overall and 90% (95% CI: 76-96) for high-grade (HG) tumours. The NPV was 98% (95% CI: 97-99) overall. The SP was 84% (95% CI: 81-86). In patients with microhaematuria, only one HG patient was missed (NPV 99%). Xpert had higher SN and NPV than cytology and UroVysion. Cytology had the highest SP (97%). In a separate SP study, Xpert had an SP of 89% in patients with benign prostate hypertrophy and 92% in prostate cancer patients. CONCLUSIONS: Xpert is an easy-to-use, noninvasive test with improved SN and NPV compared with cytology and UroVysion, representing a promising tool for identifying haematuric patients with a low likelihood of BC who might not need to undergo cystoscopy. PATIENT SUMMARY: Xpert is an easy-to-perform urine test with good performance compared with standard urine tests. It should help identify (micro)haematuria patients with a very low likelihood to have bladder cancer.

Postoperative paralytic ileus after cytoreductive surgery combined with heated intraperitoneal chemotherapy.

BACKGROUND: Patients with peritoneal malignancy treated by cytoreductive surgery (CRS) combined with hyperthermic intraperitoneal chemotherapy (HIPEC) are prone to develop postoperative paralytic ileus (POI). POI is associated with significant increase in both morbidity and mortality. CRS and HIPEC commonly result in prolonged POI (PPOI). The objective was to clarify the extent of PPOI in patients treated by CRS and HIPEC for peritoneal malignancy. METHODS: This was a prospective multicenter study including patients operated with CRS and HIPEC at the Department of Surgery, Aarhus University Hospital, Denmark and the Peritoneal Malignancy Institute, Basingstoke, United Kingdom. A total of 85 patients were included over 5 months. Patients prospectively reported parameters of postoperative gastrointestinal function in a diary from post-operative day 1 (POD1) until discharge. PPOI was defined as first defecation on POD6 or later. RESULTS: Median time to first flatus passage was 4 days (range 1-12). Median time to first defecation was 6 days (1-14). Median time to removal of nasojejunal tube was 4 days (3-13) and 7 days (1-43) for nasogastric tube. Forty-six patients (54%) developed PPOI. Patients with PPOI had longer time to first flatus (p<0.0001) and longer time to removal of nasojejunal tube (p=0.001). Duration of surgery correlated to time to first flatus (p=0.015) and time to removal of nasogastric or nasojejunal tube (p<0.0001) but not to time to first defecation (p=0.321). CONCLUSIONS: Postoperative gastrointestinal paralysis remains a common and serious problem in patients treated with CRS and HIPEC.

A novel tracer for in vivo optical imaging of fatty acid metabolism in the heart and brown adipose tissue.

Multiplexed imaging is essential for the evaluation of substrate utilization in metabolically active organs, such as the heart and brown adipose tissue (BAT), where substrate preference changes in pathophysiologic states. Optical imaging provides a useful platform because of its low cost, high throughput and intrinsic ability to perform composite readouts. However, the paucity of probes available for in vivo use has limited optical methods to image substrate metabolism. Here, we present a novel near-infrared (NIR) free fatty acid (FFA) tracer suitable for in vivo imaging of deep tissues such as the heart. Using click chemistry, Alexa Fluor 647 DIBO Alkyne was conjugated to palmitic acid. Mice injected with 0.05 nmol/g bodyweight of the conjugate (AlexaFFA) were subjected to conditions known to increase FFA uptake in the heart (fasting) and BAT [cold exposure and injection with the ß3 adrenergic agonist CL 316, 243(CL)]. Organs were subsequently imaged both ex vivo and in vivo to quantify AlexaFFA uptake. The blood kinetics of AlexaFFA followed a two-compartment model with an initial fast compartment half-life of 0.14 h and a subsequent slow compartment half-life of 5.2 h, consistent with reversible protein binding. Ex vivo fluorescence imaging after overnight cold exposure and fasting produced a significant increase in AlexaFFA uptake in the heart (58 ± 12%) and BAT (278 ± 19%) compared to warm/fed animals. In vivo imaging of the heart and BAT after exposure to CL and fasting showed a significant increase in AlexaFFA uptake in the heart (48 ± 20%) and BAT (40 ± 10%) compared to saline-injected/fed mice. We present a novel near-infrared FFA tracer, AlexaFFA, that is suitable for in vivo quantification of FFA metabolism and can be applied in the context of a low cost, high throughput, and multiplexed optical imaging platform.

Stiffness based enrichment of leukemia cells using microfluidics.

To improve the survival rate of cancer patients, new diagnosis strategies are necessary to detect lower levels of cancer cells before and after treatment regimens. The scarcity of diseased cells, particularly in residual disease after treatment, demands highly sensitive detection approaches or the ability to enrich the diseased cells in relation to normal cells. We report a label-free microfluidic approach to enrich leukemia cells from healthy cells using inherent differences in cell biophysical properties. The microfluidic device consists of a channel with an array of diagonal ridges that recurrently compress and translate flowing cells in proportion to cell stiffness. Using devices optimized for acute T cell leukemia model Jurkat, the stiffer white blood cells were translated orthogonally to the channel length, while softer leukemia cells followed hydrodynamic flow. The device enriched Jurkat leukemia cells from white blood cells with an enrichment factor of over 760. The sensitivity, specificity, and accuracy of the device were found to be > 0.8 . The values of sensitivity and specificity could be adjusted by selecting one or multiple outlets for analysis. We demonstrate that low levels of Jurkat leukemia cells (1 in 10 4 white blood cells) could be more quickly detected using flow cytometry by using the stiffness sorting pre-enrichment. In a second mode of operation, the device was implemented to sort resistive leukemia cells from both drug-sensitive leukemia cells and normal white blood cells. Therefore, microfluidic biomechanical sorting can be a useful tool to enrich leukemia cells that may improve downstream analyses.

Biologic Debris Retention in Implant Impression Copings, Scan Bodies, and Laboratory Analogs.

PURPOSE: Capturing implant position in impression-making procedures commonly involves transfer devices, such as implant impression copings and laboratory analogs. These components are intricately machined, including the lumen, and often include additional features for prevention of screw dislodgment. The Centers for Disease Control and Prevention recommends all surfaces in contact with human bodily fluid be disinfected with hospital-grade disinfectant. The ability of these components to harbor biologic contaminant material has not yet been determined, especially with regard to internal configuration, combined with the knowledge that many clinicians and laboratories use a spray disinfectant, which may limit disinfectant contact. The aim of this study was to determine the site and extent of contamination occurring on implant components following clinical impressions and laboratory procedures. MATERIALS AND METHODS: The study design included forensic staining and subsequent analysis of 60 used impression copings, 10 used laboratory analogs, and 10 new components as controls. RESULTS: Staining was found on 100% of impression copings used in vivo, indicating that biologic material had reached multiple sites on both internal and external surfaces of the components. Staining was also found on the internal aspect of used implant analogs, indicating transfer of biologic material from the impression coping and screw. None of the new control components presented staining at any site. Staining highlighted difficult areas to debride, particularly components with difficult or impossible access for cleaning and disinfection. CONCLUSION: Phloxine B staining indicated the ability of biologic material to reach all areas of the implant components. Having demonstrated the difficulty, sometimes impossibility, of accessing areas of these implant components, there is a need to develop protocols to reduce risk of potential transmission of infective material via implant components. Further study is warranted to determine the potential for transmission of infective material due to inadequate disinfection processes of implant componentry.

Redox Regulation via Glutaredoxin-1 and Protein S-Glutathionylation.

Significance: Over the past several years, oxidative post-translational modifications of protein cysteines have been recognized for their critical roles in physiology and pathophysiology. Cells have harnessed thiol modifications involving both oxidative and reductive steps for signaling and protein processing. One of these stages requires oxidation of cysteine to sulfenic acid, followed by two reduction reactions. First, glutathione (reduced glutathione [GSH]) forms a S-glutathionylated protein, and second, enzymatic or chemical reduction removes the modification. Under physiological conditions, these steps confer redox signaling and protect cysteines from irreversible oxidation. However, oxidative stress can overwhelm protein S-glutathionylation and irreversibly modify cysteine residues, disrupting redox signaling. Critical Issues: Glutaredoxins mainly catalyze the removal of protein-bound GSH and help maintain protein thiols in a highly reduced state without exerting direct antioxidant properties. Conversely, glutathione S-transferase (GST), peroxiredoxins, and occasionally glutaredoxins can also catalyze protein S-glutathionylation, thus promoting a dynamic redox environment. Recent Advances: The latest studies of glutaredoxin-1 (Glrx) transgenic or knockout mice demonstrate important distinct roles of Glrx in a variety of pathologies. Endogenous Glrx is essential to maintain normal hepatic lipid homeostasis and prevent fatty liver disease. Further, in vivo deletion of Glrx protects lungs from inflammation and bacterial pneumonia-induced damage, attenuates angiotensin II-induced cardiovascular hypertrophy, and improves ischemic limb vascularization. Meanwhile, exogenous Glrx administration can reverse pathological lung fibrosis. Future Directions: Although S-glutathionylation modifies many proteins, these studies suggest that S-glutathionylation and Glrx regulate specific pathways in vivo, and they implicate Glrx as a potential novel therapeutic target to treat diverse disease conditions. Antioxid. Redox Signal. 32, 677-700.